Journal: Blood Vessels, Thrombosis & Hemostasis

Article Title: Resilience of the bone marrow vascular architecture to aging and hematological stress

doi: 10.1016/j.bvth.2026.100156

Figure Lengend Snippet: The architecture of CD31 hi vessels in murine femora is highly conserved. (A) Femora from young adult naïve, aged, and blood-let mice were excised, benzoic acid benzyl ester-cleared, imaged by LSFM in stitched tile scans, and the resulting 3D data sets segmented for CD31 hi and CD105 + vasculature. (B-D) Quantitative analysis of the CD31 hi vasculature in 5 young adult male mice (n = 5). (B) Distribution of branch hierarchy levels with a Gaussian-like peak at level 14. (C) Branch length distribution peaking at ∼100 μm with extending to 3000 μm. (D) Branch diameter distribution ranging from 9 μm to 70 μm, peaking at 19 μm. (E-G) Quantitative analysis of the CD31 hi vasculature in 5 young adult female mice (n = 5), same as (B-D) for male mice. (H) 3D reconstruction of the CD31 hi vasculature for the young mice shown in panels B-G demonstrating high structural conservation. Scale bar, 1 mm. An overlay of all 5 male vessel trees is shown in .

Article Snippet: CD105 + vessels were segmented in Imaris and skeletonized to assess parameters such as vessel volume, vessel-vessel distance, branch length, and branching degree.

Techniques:

Journal: Blood Vessels, Thrombosis & Hemostasis

Article Title: Resilience of the bone marrow vascular architecture to aging and hematological stress

doi: 10.1016/j.bvth.2026.100156

Figure Lengend Snippet: CD105 + vessels exhibit conserved architecture and proximal narrowing in young mice. (A) Representative midshaft section of CD105 + vasculature (green) in a young naïve male mouse. Bottom row: fluorescence intensity, segmented surface, skeletonized tree, and the vessel-to-vessel distance map (200-μm section). Skeleton (cyan), end points (blue), and branching points (red). Gray outline indicates marrow boundary. Scale bars, 500 μm. (B-D) Distribution of (B) vessel diameter, (C) branching angle, and (D) vessel-to-vessel spacing in young naïve male (top) and female (bottom) mice (n = 5 per sex). See “Methods” and for detailed results. (E-J) Regional analyses along the femoral shaft divided into 9 overlapping tiles (see ). Distribution of local values of (E) vessel diameter, (F) branching angle, and (G) vessel-to-vessel distance in representative hip, mid, and knee regions of the shaft. The top row shows results of male mice, the bottom row of female mice. (H-J) Averaged values of (H) vessel diameter, (I) branching angle, and (J) vessel-to-vessel distance for each region: hip-proximal (yellow), midshaft (magenta), and knee-proximal (grape). Filled bars represent male mice, lighter colored shaded bars female mice. Shown are mean ± SEM. SEM, standard error of the mean.

Article Snippet: CD105 + vessels were segmented in Imaris and skeletonized to assess parameters such as vessel volume, vessel-vessel distance, branch length, and branching degree.

Techniques: Fluorescence

Journal: Blood Vessels, Thrombosis & Hemostasis

Article Title: Resilience of the bone marrow vascular architecture to aging and hematological stress

doi: 10.1016/j.bvth.2026.100156

Figure Lengend Snippet: Morphological differences in CD105 + small vessels of aged vs young mice. (A-B) Representative midshaft femur sections from an aged male (A) and a young, blood-let mouse (B), showing segmented CD105 + surfaces and skeletonized vasculature. Color coding as in . Scale bar, 500 μm. (C-E) Distribution of vessel diameters along the femoral shaft. (F-H) Mean vessel diameter per region along the femur. (I-K) Distribution of vessel-to-vessel distances. (L-N) Regional mean vessel-to-vessel distances. Dashed lines and lighter shaded bars indicate female data. See for further details.

Article Snippet: CD105 + vessels were segmented in Imaris and skeletonized to assess parameters such as vessel volume, vessel-vessel distance, branch length, and branching degree.

Techniques:

Journal: Blood Vessels, Thrombosis & Hemostasis

Article Title: Resilience of the bone marrow vascular architecture to aging and hematological stress

doi: 10.1016/j.bvth.2026.100156

Figure Lengend Snippet: Regional changes in the CD105 + microvascular complexity along the femur. (A) Fraction of long branches (>100 μm) in young naïve (blue), blood-let (red), and aged (gray) mice (filled symbols, male; open symbols, females). (B) Proportion of vessels running parallel to the central vein. (C) Fraction of vessels with diameters <25 μm. (D) Ratio of vessel segments to branching points. (E) Ratio of end points to branching points. (F) Fraction of branching points with ≥4 outgoing branches, only branches longer than 25 μm and branching degree ≥3 were included. All data shown as mean ± SEM. SEM, standard error of the mean.

Article Snippet: CD105 + vessels were segmented in Imaris and skeletonized to assess parameters such as vessel volume, vessel-vessel distance, branch length, and branching degree.

Techniques:

Journal: Blood Vessels, Thrombosis & Hemostasis

Article Title: Resilience of the bone marrow vascular architecture to aging and hematological stress

doi: 10.1016/j.bvth.2026.100156

Figure Lengend Snippet: Modest increased MK volume in aged male mice. (A) Representative midshaft section showing CD105 + MKs (magenta) in a young adult male mouse. Bottom row: fluorescence intensity, segmented surface, MK outlines, and MK-vessel distance map (200-μm section). MK outlines and distance transform shown for a single slice. Gray outline indicates marrow boundary. Scale bar, 500 μm. (B-D) Quantification of MK properties in young (blue), aged (gray), and blood-let (red) mice (filled symbols: males; open symbols: females). (B) MK-to-AF volume ratio; (C) MK surface-to-volume ratio; and (D) proportion of MK surface within 5 μm of a vessel. (E-G) MK-vessel distance distributions; dashed lines: female mice. (H-J) Regional mean MK-vessel distances for hip, mid, and knee. Lighter colored shaded bars: females. All data shown as mean ± SEM. See “Methods” and and for details. BM, bone marrow; SEM, standard error of the mean; Sur./Vol., surface/volume.

Article Snippet: CD105 + vessels were segmented in Imaris and skeletonized to assess parameters such as vessel volume, vessel-vessel distance, branch length, and branching degree.

Techniques: Fluorescence

Journal: bioRxiv

Article Title: PS FAD mutants and γ-secretase inhibition accumulate VEGFR2-derived peptide VCTF1 suppressing brain VEGFR2 dimerization, angiogenesis and neuroprotection

doi: 10.64898/2026.05.12.724648

Figure Lengend Snippet: (A) , Wild-type (WT) and KI mice HTRZ for either PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T) were infused with vehicle (0.2% BSA in PBS) or VEGF-A (a total of 3.5μg in 100μl of vehicle) through the carotid artery for 15 days using a mini osmotic pump as in Methods. Left: Brain coronal sections (40μm thick) were prepared and immunostained with anti-Col IV antibodies to visualize brain vessels. Enhanced visualization surfaces were generated using Imaris software from representative confocal images of ipsilateral hemispheres. Scale bar: 50μm. Right: Graph shows total vessel length density in WT and PS1 FAD brains quantified using Imaris 9.9 software as in Methods. (B) , WT and HTRZ for PS1 FAD mutants M146V or I213T mice were injected through the carotid artery for 20 minutes with either vehicle or 100ng of VEGF-A in vehicle prepared as in 1A using a catheter as described in Methods. Brain microvessels (MV) were isolated as in Methods, lysed in Triton X-100 buffer, and subjected to immunoprecipitation (IP) with anti-VEGFR2 antibody or control IgG. Left: IPs were analyzed on Western blots (WBs) using anti-endoglin or anti-VEGFR2 antibodies (upper panel). Input samples are shown in lower panel. β-actin: loading control. Right: Graph shows quantification of endoglin co-IPed with VEGFR2, normalized to IPed VEGFR2. (C) , WT mice were infused for 15 days through the carotid artery with vehicle or VEGF-A in vehicle as in 1A using a mini osmotic pump (as in 1A). For RO injection, mice were treated with vehicle (2% DMSO, 30% PEG 300, 5% Tween-80 in ddH2O) or RO in vehicle (5mg/kg body weight) via five injections in tail vein one injection every three days, with first injection administered 1 hour before osmotic pump implantation. Brain coronal sections (40μm) were prepared and immunostained with anti-Col IV antibodies as in 1A. Left: Representative confocal images of ipsilateral hemispheres are shown prepared as in 1A. Scale bar: 50μm. Right: Graph shows total vessel length density quantified using Imaris software as in 1A. (D) , WT adult mice were treated with either 50μl vehicle as in 1C or 1mg/kg RO in vehicle via carotid artery as in Methods. 15-16 hrs later, 50 μl vehicle prepared as in 1A or 100ng VEGF-A in vehicle was administered via carotid artery for 10-20 minutes using a catheter as in 1B. Brain MVs were isolated and extracted as in 1B. Left: p-VEGFR2 (Tyr1054/Tyr1059), VEGFR2, p-ERK1/2 and ERK1/2 are detected on WBs of extracts with specific antibodies in MV extracts. Vinculin: loading control. Right: Graphs show fold change of phosphorylated to total protein ratio. (E) , WT mice and mice HTRZ for PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T) were treated with vehicle or VEGF-A via carotid artery for 10-20 minutes using a catheter as in 1D. Brain MVs were isolated and extracted as in 1B. Left: p-VEGFR2 (Tyr1054/Tyr1059), VEGFR2, p-ERK1/2 and ERK1/2 are detected on WBs of extracts with specific antibodies in MV extracts. β-actin: loading control. Right: Graphs show fold change of phosphorylated to total protein ratio. (F) , WT pCECs were prepared and treated as in Methods with vehicle (DMSO) or RO (200nM in DMSO) and then stimulated with either vehicle (PBS) or VEGF-A (20ng in PBS) for 15min. Upper: Cells were co-immunostained with either anti-VEGFR2 antibodies (green) or early endosome marker Rab5 (red) and cell nuclei were stained with Hoechst (blue) as in Methods. Yellow fluorescence in merged images indicates co-localization of VEGFR2 with Rab5. Scale bar 0.5μm. Lower: Graph shows percent of VEGFR2 co-localized with Rab5 in RO-treated WT cells compared to vehicle-treated cells measured with Imaris software. (G) , pCECs from either WT or mice HTRZ for PS1 FAD mutant M146V (WT/M146V) or I213T (WT/I213T), were stimulated with vehicle or VEGF-A in vehicle as in 1F. Upper: Cells were co-stained with anti-VEGFR2 antibodies and early endosome marker Rab5 as in 1F. Cell nuclei were stained with Hoechst (blue) as in 1F. Yellow fluorescence in merged images indicates co-localization of VEGFR2 with Rab5. Scale bar 0.5μm. Lower: Graph shows percent of VEGFR2 co-localized with Rab5 in PS1 FAD WT/M146V or WT/I213T HTRZ mice compared to WT measured with Imaris software. For Figs A-G, data are shown as Mean ± S.E. from at least three independent experiments or as indicated in the dot plots. Statistical analysis was performed using two-way ANOVA followed by Tukey post-hoc test. ns = not significant, *p<0.05, **p<0.01, ***p<0.001.

Article Snippet: Mouse monoclonal anti-Flag tag (M2; F1804) was from Millipore Sigma, anti-GAPDH (2118S) from Cell Signaling Technologies (Beverly, MA), anti-VEGFR2 (OTI12C1) from Origene, anti-endoglin (CD-105; NBP2-22122) and anti-LAMP2 (NBP2-22217) from Novus Biologicals, Inc, anti-Rab5 (D-11) and anti-Rab7 (B-3) from Santa Cruz Biotechnology, Inc. Chicken polyclonal anti-GFAP (ab4674) was from Abcam.

Techniques: Mutagenesis, Generated, Software, Injection, Isolation, Immunoprecipitation, Control, Western Blot, Marker, Staining, Fluorescence

Journal: bioRxiv

Article Title: PS FAD mutants and γ-secretase inhibition accumulate VEGFR2-derived peptide VCTF1 suppressing brain VEGFR2 dimerization, angiogenesis and neuroprotection

doi: 10.64898/2026.05.12.724648

Figure Lengend Snippet: (A), Brain tissue extracts were prepared as in Methods from twelve PS1 FAD patients each carrying a different PS1 mutation, and twelve non-demented controls described in Methods. Left: VEGFR2 dimers and monomers were detected in brain extracts on WBs using anti-VEGFR2 antibody D5B1. Representative gels with control (C1-4) or FAD samples (FAD1-4) expressing mutants P264L, A260V, N135S and P242H respectively are shown. Vinculin: loading control. Right: Graph shows the fold change in VEGFR2 dimer to monomer ratio of FAD and control samples. (B), Brain tissue extract from control and PS1 FAD patient brains described in 7A were prepared and IPed with anti-endoglin antibody (ab252345) or IgG as in Methods. Upper panel: VEGFR2 co-IPed with endoglin was detected on WBs using an anti-VEGFR2 antibody as in 7A. Lower panel: Input samples are shown. Representative gel with control samples (C1, C2) and FAD samples (FAD1, FAD2) expressing mutants A260V and P264L respectively is shown. β-actin: loading control. Right: Graph shows relative levels of VEGFR2 co-precipitated with endoglin. (C), Brain sections from control and PS1 FAD patients were prepared as in Methods and stained for Col IV as in 1A. Upper: Representative images show brain vessels in either PS1 FAD or control (CT) brain sections. Scale bar: 80μm. Lower: Graph shows total vessel length density in PS1 FAD and CT brains quantified with Imaris software as in 1A. A-C , bars represent Mean ± S.E. For statistical analysis, unpaired t-test was performed. *p < 0.05, **p<0.01 and ***p<0.001.

Article Snippet: Mouse monoclonal anti-Flag tag (M2; F1804) was from Millipore Sigma, anti-GAPDH (2118S) from Cell Signaling Technologies (Beverly, MA), anti-VEGFR2 (OTI12C1) from Origene, anti-endoglin (CD-105; NBP2-22122) and anti-LAMP2 (NBP2-22217) from Novus Biologicals, Inc, anti-Rab5 (D-11) and anti-Rab7 (B-3) from Santa Cruz Biotechnology, Inc. Chicken polyclonal anti-GFAP (ab4674) was from Abcam.

Techniques: Mutagenesis, Control, Expressing, Staining, Software

Journal: Cells

Article Title: Alterations in Circulating Progenitor Cell Composition in Rheumatoid Arthritis

doi: 10.3390/cells15080726

Figure Lengend Snippet: CD235a + and CD45 − CD31 − stromal cells circulating in peripheral blood. ( a ) CD45/CD31 dot plot showing populations 16, 17, 18, 19 and 20. ( b ) UMAP shows the size of each cluster, and ( c ) TriMAP shows the relative cluster location. CD45/CD31 plots highlighting the expression of ( d ) CD235a, ( e ) CD133, ( f ) CD34 and ( g ) CD105. The intensity scale indicates high expression of the marker in red. Frequency of ( h ) Pop 20, ( i ) Pop 17, and ( j ) Pop 18. Mann-Whitney test. Dot plots showing the expression of ( k ) CD34/CD133, ( l ) CD271/ CD105, and CD90 with ( m ) CD105, ( n ) CD271, and ( o ) PDPN in Pop 16 and Pop 19. Frequency of ( p ) Pop 16 and ( q ) Pop 19. t -test for Pop 16; Mann–Whitney test for Pop 19. All dot plots show mean and SD, n = 12 RA and 9 HC.

Article Snippet: CD105 , VioBlue-A , 130-099-666 , Miltenyi Biotec , 1/100 , V3.

Techniques: Expressing, Marker, MANN-WHITNEY